Although humic acids (HAs) from peat exhibit various therapeutic properties, there is little information available concerning their physicochemical and antioxidant properties. To address this issue, nine different types of peat, including oligotrophic, mesotrophic, and minerotrophic peat samples, were used for isolation of HA fractions by basic (HAb) and pyrophosphate (HAp) extractions. Physical parameters of the HAs were analyzed by UV-Vis, fluorescent, infrared (IR), and electron paramagnetic resonance (EPR) spectroscopy. Average Mr of the fractions ranged from 17.2 to 39.7 kDa, while their humification index (HIX) varied from 0.49 to 1.21. HAp fractions had a higher content of aromatic structures compared to HAb fractions. Moreover, HAp fractions had a significantly higher content of phenolic OH groups (3.6 ± 0.5 mmol/g) versus HAb (3.1 ± 0.5 mmol/g). All HA fractions exhibited antioxidant activity in radical scavenging and electrochemical assays, and their EPR signal had a single line with g = 2.0035, which is consistent with semiquinone type radicals. Furthermore, the HIX was found to be important in determining the number of semiquinone-type free radicals in the HA structures. Overall, these data provide a molecular basis to explain at least part of the beneficial therapeutic properties of peat-derived HAs.

Humic substances (HS) are natural supramolecular systems of high- and low-molecular-weight compounds with distinct immunomodulatory and protective properties. The key beneficial biological activity of HS is their antioxidant activity. However, systematic studies of the antioxidant activity of HS against biologically relevant peroxyl radicals are still scarce. The main objective of this work was to estimate the antioxidant capacity (AOC) of a broad set of HS widely differing in structure using an oxygen radical absorption capacity (ORAC) assay. For this purpose, 25 samples of soil, peat, coal, and aquatic HS and humic-like substances were characterized using elemental analysis and quantitative 13C solution-state NMR. The Folin–Ciocalteu method was used to quantify total phenol (TP) content in HS. The determined AOC values varied in the range of 0.31–2.56 μmol Trolox eqv. mg−1, which is close to the values for ascorbic acid and vitamin E. Forward stepwise regression was used to reveal the four main factors contributing to the AOC value of HS: atomic C/N ratio, content of O-substituted methine and methoxyl groups, and TP. The results obtained clearly demonstrate the dependence of the AOC of HS on both phenolic and non-phenolic moieties in their structure, including carbohydrate fragments.

We studied activation of macrophages with humic acids extracted from peat of large deposits in the Tomsk region by two extraction methods: by hydroxide or sodium pyrophosphate. Humic acid of lowland peat types containing large amounts of aromatic carbon, phenolic and alcohol groups, carbohydrate residues and ethers, irrespectively of the extraction methods contained LPS admixture that probably determines their activating properties. Humic acid of upland peat types characterized by high content of carbonyl, carboxyl, and ester groups enhance NO production and reduce arginase expression, but these effects were minimized when sodium hydroxide was used as an extraction solvent. Pyrophosphate samples of the upland peat types were characterized by aromaticity and diversity of functional groups and have a significant advantage because of they induce specific endotoxin-independent stimulating action on antigen presenting cells.

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The active ingredient extracted from the peat humic substances was characterized by physicochemical parameters evaluated by UV- and IR-spectroscopy, titration, and elemental (C, H, N) analysis. The cardiovascular effects of this ingredient were examined on isolated Langendorff-perfused rat heart. It was found that the active substance in a concentration range of 0.01-0.1 mg/ml produced a vasodilating effect; in addition, it decreased the end-diastolic and left-ventricular developed pressures.